• Title/Summary/Keyword: biochemical property

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Some Observations on the Organelles Participating in the Biliary Excretion in the Hepatocyte of the Biligrafin Injected Mouse (Biligrafin 투여 마우스 간세포의 미세구조적 및 세포화학적 연구)

  • Kim, Hyang;Shin, Young-Chul
    • Applied Microscopy
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    • v.23 no.2
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    • pp.53-77
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    • 1993
  • In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

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Characterization of Bacillus licheniformis SCK A08 with Antagonistic Property Against Bacillus cereus and Degrading Capacity of Biogenic Amines (Bacillus cereus에 대한 길항적 저해 작용과 biogenic amines 분해 능력을 지닌 Bacillus licheniformis SCK A08 균의 특성)

  • Lee, Eon Sil;Kim, Yong Sang;Ryu, Myeong Seon;Jeong, Do Yeon;Uhm, Tai Boong;Cho, Sung Ho
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.40-46
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    • 2014
  • We have screened Bacillus strains suitable for the fermentation of soybean products with respect to the control of Bacillus cereus and the reduction of biogenic amines. Of 26 isolates, a strain named as the SCK A08 carried antimicrobial activity against B. cereus and Staphylococcus aureus, major food poisoning species in soybean products. PCR analysis revealed that the SCK A08 strain did not contain genes for Bacillus cereus toxins including nonhemolytic enterotoxin, hemolytic enterotoxin, cytotoxin K, cereulide and certrax. The SCK A08 strain could degrade histamine, tyramine, putrescine, and cadaverine by 67.41%, 76.59%, 57.32%, and 50.69%, respectively, during fermentation in cooked soybeans containing 0.5% (w/w) of each biogenic amine. The morphological and biochemical properties and phylogenetic relationships based on 16S rRNA gene sequences indicated that the isolate was most closely related to Bacillus licheniformis. Use of the strain SCK A08 would be a potential measure to overcome two hygienic problems that were frequently faced during manufacture of traditionally fermented soybean products.

Identification and Characterization of Microbial Community in the Coelomic Fluid of Earthworm (Aporrectodea molleri)

  • Yakkou, Lamia;Houida, Sofia;Dominguez, Jorge;Raouane, Mohammed;Amghar, Souad;Harti, Abdellatif El
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.391-402
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    • 2021
  • Earthworms play an important role in soil fertilization, interacting continually with microorganisms. This study aims to demonstrate the existence of beneficial microorganisms living in the earthworm's immune system, the coelomic fluid. To achieve this goal, a molecular identification technique was performed, using cytochrome c oxidase I (COI) barcoding to identify abundant endogenic earthworms inhabiting the temperate zone of Rabat, Morocco. Then, 16S rDNA and ITS sequencing techniques were adopted for bacteria and fungi, respectively. Biochemical analysis, showed the ability of bacteria to produce characteristic enzymes and utilize substrates. Qualitative screening of plant growth-promoting traits, including nitrogen fixation, phosphate and potassium solubilization, and indole acetic acid (IAA) production, was also performed. The result of mitochondrial COI barcoding allowed the identification of the earthworm species Aporrectodea molleri. Phenotypic and genotypic studies of the sixteen isolated bacteria and the two isolated fungi showed that they belong to the Pseudomonas, Aeromonas, Bacillus, Buttiauxella, Enterobacter, Pantoea, and Raoultella, and the Penicillium genera, respectively. Most of the isolated bacteria in the coelomic fluid showed the ability to produce β-glucosidase, β-glucosaminidase, Glutamyl-β-naphthylamidase, and aminopeptidase enzymes, utilizing substrates like aliphatic thiol, sorbitol, and fatty acid ester. Furthermore, three bacteria were able to fix nitrogen, solubilize phosphate and potassium, and produce IAA. This initial study demonstrated that despite the immune property of earthworms' coelomic fluid, it harbors beneficial microorganisms. Thus, the presence of resistant microorganisms in the earthworm's immune system highlights a possible selection process at the coelomic fluid level.

Anti-fatigue effect of tormentic acid through alleviating oxidative stress and energy metabolism-modulating property in C2C12 cells and animal models

  • Ho-Geun Kang;Jin-Ho Lim;Hee-Yun Kim;Hyunyong Kim;Hyung-Min Kim;Hyun-Ja Jeong
    • Nutrition Research and Practice
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    • v.17 no.4
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    • pp.670-681
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    • 2023
  • BACKGROUND/OBJECTIVES: Oxidative stress is caused by reactive oxygen species and free radicals that accelerate inflammatory responses and exacerbate fatigue. Tormentic acid (TA) has antioxidant and anti-inflammatory properties. Thus, the aim of present study is to determine the fatigue-regulatory effects of TA in H2O2-stimulated myoblast cell line, C2C12 cells and treadmill stress test (TST) and forced swimming test (FST) animal models. MATERIALS/METHODS: In the in vitro study, C2C12 cells were pretreated with TA before stimulation with H2O2. Then, malondialdehyde (MDA), lactate dehydrogenase (LDH), creatine kinase (CK) activity, tumor necrosis factor (TNF)-α, interleukin (IL)-6, superoxide dismutase (SOD), catalase (CAT), glycogen, and cell viability were analyzed. In the in vivo study, the ICR male mice were administered TA or distilled water orally daily for 28 days. FST and TST were then performed on the last day. In addition, biochemical analysis of the serum, muscle, and liver was performed. RESULTS: TA dose-dependently alleviated the levels of MDA, LDH, CK activity, TNF-α, and IL-6 in H2O2-stimulated C2C12 cells without affecting the cytotoxicity. TA increased the SOD and CAT activities and the glycogen levels in H2O2-stimulated C2C12 cells. In TST and FST animal models, TA decreased the FST immobility time significantly while increasing the TST exhaustion time without weight fluctuations. The in vivo studies showed that the levels of SOD, CAT, citrate synthase, glycogen, and free fatty acid were increased by TA administration, whereas TA significantly reduced the levels of glucose, MDA, LDH, lactate, CK, inflammatory cytokines, alanine transaminase, aspartate transaminase, blood urea nitrogen, and cortisol compared to the control group. CONCLUSIONS: TA improves fatigue by modulating oxidative stress and energy metabolism in C2C12 cells and animal models. Therefore, we suggest that TA can be a powerful substance in healthy functional foods and therapeutics to improve fatigue.

Recent Progress in Colorimetric Assays Using the Absorption of Plasmonic Gold Nanoparticles (플라즈모닉 금 나노입자의 흡광 특성을 활용한 생화학적 비색 분석법 연구 동향)

  • Bong-Geun Kim;Sang Bin Yoon;Sukyeong Hwang;Hyon Bin Na
    • Applied Chemistry for Engineering
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    • v.35 no.2
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    • pp.67-78
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    • 2024
  • Light absorption has potential as a signal in biochemical analyses due to its simplicity in measurement and interpretational clarity. Among substances that generate absorption signals, gold nanoparticles possess advantages such as chemical stability, biological compatibility, and unique optical properties from the localized surface plasmon resonance (LSPR) in the visible light range. They also exhibit versatility compared to other colorimetric substances effective only for specific target molecules, as they easily conjugate with various detection active substances like antibodies and aptamers. Particularly due to advantages such as low cost, ease of particle synthesis, and high environmental stability compared to enzyme-based colorimetric methods, gold nanoparticles are extensively researched as signal substances in colorimetric assays. This review summarizes various strategies utilizing gold nanoparticles as absorption signal substances, focusing on recent research. Based on the characteristics of gold nanoparticles, where the optical property is influenced by particle morphology, literature is classified and reviewed based on strategies controlling the shape of gold nanoparticles during signal generation. Through this, it is observed that gold nanoparticles, which have been used as absorption signal substances, continue to be actively researched, affirming their potential for broad and continuous improvement in the future.

The Physio-ecological Characteristics of Golden Apple Snails (Pomacea canaliculata) and the Cause of Their Massive Death used for Weed Control in Wet Rice Paddies (왕우렁이(Pomacea canaliculata)의 생리·생태학적 특성 및 논 잡초방제용으로 투입된 왕우렁이의 집단패사 원인)

  • Lee, Sang-Beom;Lee, Sang-Min;Ko, Byong-Gu;Lee, Cho-Rong;Kim, Jin-Ho
    • Korean Journal of Organic Agriculture
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    • v.26 no.2
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    • pp.297-316
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    • 2018
  • The golden apple snail (GAS, Pomacea canaliculata) is an invasive freshwater snail. The GAS was introduced in Korea without prior studies on the possibility of crop damage or its impact on the natural ecosystem. The freshwater apple snails can be found typically in ponds, rice paddies, irrigation canals, roadside ditches or slower portions of streams. In this study, we were carried out to investigate the assessment of physiological and ecological characteristics, environmental characteristics inhabited area in winter season and cause of massive death at one time of golden apple snails used for weed control in wet rice paddies. The GAS was introduced from Japan to Korea for commercial production as a dietary protein supplement. The golden apple snail was also used a recently for weed control in wet rice cultivation. The species of freshwater Pomacea snails is belonging to the genus Pomacea, family ampulariidae, order mesogastropoda, subclass pulmonata, class gastropoda, phylum mollusca. The GAS spread into irrigation ditches and natural waterways. It is now distributed in ponds and canals near rice fields of southern parts of the country and has overwintered. It increases its cold hardiness before winter. However, the physiological mechanism of cold hardiness in molluscs is poorly understood, especially in freshwater molluscs. Our results on physio-ecological characteristics of the Pomacea apple snail showed that the ratio of males to females was 1: 1.99~2.33. The daily growth was 87.7 mg in weight, 0.31 mm in height and 0.33 mm in width of the their shell. On the other hand, the golden apple snails were very high to resistance on drying condition and survived rate about 80% up to 3 months. The inhabitation of GAS was no statistical significant impacts on the water quality. An important property of aqueous solutions is agricultural water quality because it affects chemical and biochemical properties such as chemical reactions, equilibrium conditions, and biological toxicity. The death rate of weed control apple snails by Ostracoda (Stenocypris hislopi) was only 2.86% and 5.71% depending on the density. Therefore, GAS was not a direct death caused by Ostracoda (Stenocypris hislopi).

Development of Benthic Macroinvertebrates Index (BMI) for Biological Assessment on Stream Environment (하천환경의 생물학적 평가를 위한 저서동물지수(BMI)의 개발)

  • Kong, Dongsoo;Son, Se-Hwan;Hwang, Soon-Jin;Won, Doo Hee;Kim, Myoung Chul;Park, Jung Ho;Jeon, Te Su;Lee, Jong Eun;Kim, Jong Hyun;Kim, Jong Sun;Park, Jaeheung;Kwak, Inn Sil;Ham, Sun Ah;Jun, Yung-Chul;Park, Young-Seuk;Lee, Jae-Kwan;Lee, Su-Woong;Park, Chang-Hee;Moon, Jeong-Suk;Kim, Jin-Young;Park, Hae Kyung;Park, Sun Jin;Kwon, Yongju;Kim, Piljae;Kim, Ah Reum
    • Journal of Korean Society on Water Environment
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    • v.34 no.2
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    • pp.183-201
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    • 2018
  • The tolerance of Korean benthic macroinvertebrates to organic pollution has been analyzed since the early 1990s. However, considering the fact that there have been related studies carried out in some European countries since the early 20th century, the history of the research in Korea is very short and there is still much knowledge to supplement. We revised the saprobic valency, the saprobic value and the indicator weight value of 190 benthic macroinvertebrates taxa through the data of water quality and individual abundance collected from 7,086 sampling units in Korea from 2008 to 2014. The individual abundance of Uracanthella (Ephemeroptera) as a representative, one of the most common and abundant taxa in Korea, showed a typical lognormal distribution to 5-day biochemical oxygen demand (BOD5) concentration, and a normal distribution to the class interval of BOD5 concentration according to saprobic series. The value combining the mean individual abundance and the relative frequency of occurrence was a more efficient indicator value than that of each property alone. Benthic Macroinertebrates Index (BMI) was newly proposed as a modification of the saprobic index of Zelinka and Marvan (1961). BMI showed extremely significant correlation (determination coefficient $r^2$ > 0.6, n = 569 sites) with the concentration of BOD5, and the coefficient was a little higher than those of the previous indices. Until now, there has been very little research on the assessment of biological integrity of benthic macroinvertebrates community in Korea. While continuing researches into improve the reliability of BMI, it is necessary to develop multimetric indices for evaluating the integrity, including the composition of species and functional guilds, and the richness and diversity of the community.

Biochemical and Physiological Properties of Fermented Skate (홍어 숙성과 기능성)

  • 최명락;유은정;임현수;박재원
    • Journal of Life Science
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    • v.13 no.5
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    • pp.675-683
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    • 2003
  • This study was conducted to investigate the physiological properties of various parts of skate body after fermentation by measuring compositional properties including pH and $NH_4^+$. Other functional properties, such as antibacterial activities, antioxidative activities and anticancer activities were measured. Effects of fermentation temperature (4, 10, $20^{\circ}C$) did not affect compositional properties of fermented skate. The pH of fermented skate extract at 4$^{\circ}C$ did not increase as much as that at 10 and $20^{\circ}C$, Particularly at $10^{\circ}C$, the pH increased rapidly after Day 1 and remained unchanged until another increase at Day 5. At 2$0^{\circ}C$, the pH increased rather rapidly at early stage of fermentation and reached 8.9 at Day 4. The pattern of $NH_4^+$ concentration of fermented skate extract was similar to that of pH. Particularly at $4^{\circ}C$ fermentation, $NH_4^+$ concentration was not affected by fermentation time. The concentration of $NH_4^+$. reached approximately $10.2\mug/mL$at $10^{\circ}C$ for Day 5 and $20^{\circ}C$ for Day 4-5, indicating the early stage of fermentation. According to physiological activities of hot water extracts of skate fillet and viscera as affected by fermentation time, antibacterial activity of 2% viscera extract concentration was 43.3% at Day 8, while there was no antibacterial activity from fillet extract. As for the antioxidative activity, fillet extract and viscera extract both at 2% concentration at Day 0 showed 61.2% and 54.4%, respectively. Anticancer activities were highest (52.7% for fillet extract and 58.3% for viscera extract) at $1,000 \mug/mL$ concentration at Day 8. Antibacterial activities and anticancer effects were relatively high as fermentation was progressed, while antioxidative activities were highest before fermentation started. As for the physiological activities of hot water extract from brain and cartilage, antibacterial activities were observed at 41.0% when 2% brain extract was added at 4 hours of incubation, while 35.8% with 2% cartilage extract at 14 hours of incubation. Antioxidative activities were not found in brain extract, but cartilage extract at 2% showed 25.0% of antioxidative activity, which was lower than fillet and viscera extract.

A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.